Cottrell J.R.,Galenea Corporation |
Levenson J.M.,Galenea Corporation |
Kim S.H.,New York Medical College |
Kim S.H.,Kyung Hee University |
And 14 more authors.
Journal of Neuroscience | Year: 2013
Working memory is an essential component of higher cognitive function, and its impairment is a core symptom of multiple CNS disorders, including schizophrenia. Neuronal mechanisms supporting working memory under normal conditions have been described and include persistent, high-frequency activity of prefrontal cortical neurons. However, little is known about the molecular and cellular basis of working memory dysfunction in the context of neuropsychiatric disorders. To elucidate synaptic and neuronal mechanisms of working memory dysfunction, we have performed a comprehensive analysis of a mouse model of schizophrenia, the forebrain-specific calcineurin knock-out mouse. Biochemical analyses of cortical tissue from these mice revealed a pronounced hyperphosphorylation of synaptic vesicle cycling proteins known to be necessary for high-frequency synaptic transmission. Examination of the synaptic vesicle cycle in calcineurin-deficient neurons demonstrated an impairment of vesicle release enhancement during periods of intense stimulation. Moreover, brain slice and in vivo electrophysiological analyses showed that loss of calcineurin leads to a gene dose-dependent disruption of high-frequency synaptic transmission and network activity in the PFC, correlating with selective working memory impairment. Finally, we showed that levels of dynamin I, a key presynaptic protein and calcineurin substrate, are significantly reduced in prefrontal cortical samples from schizophrenia patients, extending the disease relevance of our findings. Our data provide support for a model in which impaired synaptic vesicle cycling represents a critical node for disease pathologies underlying the cognitive deficits in schizophrenia. © 2013 the authors.
Larson K.C.,Galenea Corporation |
Draper M.P.,Paratek Pharmaceuticals Inc. |
Lipko M.,Nencki Institute of Experimental Biology |
Dabrowski M.,Nencki Institute of Experimental Biology
Inflammation Research | Year: 2010
Background: Inflammation plays a central role in many neurodegenerative diseases, including Parkinson's, Alzheimer's, multiple sclerosis, amyotrophic lateral sclerosis, and AIDS dementia. Microglia are the resident macrophages of the central nervous system and are the cells primarily responsible for the inflammatory component of these diseases. Methods: Using gene expression profiling, we compared the profile of the neurospecific microglial cell line BV-2 after LPS stimulation to that of a macrophage cell line (J774A.1) stimulated with LPS. Results: A set of 77 genes that were modulated only in microglial cells after LPS stimulation was identified. One gene of interest, Gng12, was investigated further to determine its ability to modify the inflammatory response. Specifically, Gng12 mRNA levels were transiently increased after LPS stimulation. In addition, overall levels of Gng12 mRNA after LPS stimulation were significantly higher in BV-2 cells as compared to macrophage cells. Conclusion: Modulating Gng12 mRNA levels using RNAi revealed a novel role for the factor in the negative regulation of the overall inflammatory response as based on effects on nitrite and TNFα levels. These data suggest that Gng12 is a negative regulator of the LPS response and may be an important factor in the overall inflammatory signaling cascade. © 2009 Birkhäuser Verlag, Basel/Switzerland.
Jensen P.C.,Copenhagen University |
Thiele S.,Copenhagen University |
Steen A.,Copenhagen University |
Elder A.,Millennium Pharmaceuticals Inc. |
And 6 more authors.
British Journal of Pharmacology | Year: 2012
Background and Purpose The majority of small molecule compounds targeting chemokine receptors share a similar pharmacophore with a centrally located aliphatic positive charge and flanking aromatic moieties. Here we describe a novel piperidine-based compound with structural similarity to previously described CCR8-specific agonists, but containing a unique phenyl-tetrazol moiety which, in addition to activity at CCR8 was also active at CCR1. Experimental Approach Single point mutations were introduced in CCR1 and CCR8, and their effect on small molecule ligand-induced receptor activation was examined through inositol trisphosphate (IP 3) accumulation. The molecular interaction profile of the agonist was verified by molecular modeling. Key Results The chemokine receptor conserved glutamic acid in TM-VII served as a common anchor for the positively charged amine in the piperidine ring. However, whereas the phenyl-tetrazol group interacted with TyrIV:24 (Tyr 172) and TyrIII:09 (Tyr 114) in the major binding pocket (delimited by TM-III to VII) of CCR8, it also interacted with TrpII:20 (Trp 90) and LysII:24 (Lys 94) in the minor counterpart (delimited TM-I to III, plus TM-VII) in CCR1. A straightening of TM-II by Ala-substitution of ProII:18 confirmed its unique role in CCR1. The extracellular loop 2 (ECL-2) contributed directly to the small molecule binding site in CCR1, whereas it contributed to efficacy, but not potency in CCR8. Conclusion and Implications Despite high ligand potency and efficacy and receptor similarity, this dual-active and bitopic compound binds oppositely in CCR1 and CCR8 with different roles of ECL-2, thereby expanding and diversifying the influence of extracellular receptor regions in drug action. © 2011 The British Pharmacological Society.
Hempel C.M.,Galenea Corporation |
Sivula M.,Galenea Corporation |
Levenson J.M.,Galenea Corporation |
Rose D.M.,Galenea Corporation |
And 6 more authors.
PLoS ONE | Year: 2011
Unbiased, high-throughput screening has proven invaluable for dissecting complex biological processes. Application of this general approach to synaptic function would have a major impact on neuroscience research and drug discovery. However, existing techniques for studying synaptic physiology are labor intensive and low-throughput. Here, we describe a new high-throughput technology for performing assays of synaptic function in primary neurons cultured in microtiter plates. We show that this system can perform 96 synaptic vesicle cycling assays in parallel with high sensitivity, precision, uniformity, and reproducibility and can detect modulators of presynaptic function. By screening libraries of pharmacologically defined compounds on rat forebrain cultures, we have used this system to identify novel effects of compounds on specific aspects of presynaptic function. As a system for unbiased compound as well as genomic screening, this technology has significant applications for basic neuroscience research and for the discovery of novel, mechanism-based treatments for central nervous system disorders. © 2011 Hempel et al.
PubMed | Yale University, Kyung Hee University, Galenea Corporation and New York Medical College
Type: Journal Article | Journal: The Journal of biological chemistry | Year: 2016
Variation in PPP3CC, the gene that encodes the isoform of the calcineurin catalytic subunit, has been reported to be associated with schizophrenia. Because of its low expression level in most tissues, there has been little research devoted to the specific function of the calcineurin A (CNA) versus the calcineurin A (CNA) and calcineurin A (CNA) catalytic isoforms. Consequently, we have a limited understanding of the role of altered CNA function in psychiatric disease. In this study, we demonstrate that CNA is present in the rodent and human brain and dephosphorylates a presynaptic substrate of calcineurin. Through a combination of immunocytochemistry and immuno-EM, we further show that CNA is localized to presynaptic terminals in hippocampal neurons. Critically, we demonstrate that RNAi-mediated knockdown of CNA leads to a disruption of synaptic vesicle cycling in cultured rat hippocampal neurons. These data indicate that CNA regulates a critical aspect of synaptic vesicle cycling and suggest that variation in PPP3CC may contribute to psychiatric disease by altering presynaptic function.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 699.79K | Year: 2011
DESCRIPTION (provided by applicant): This application, Novel serotonergic, pro-cognitive antipsychotic therapies , is in response to PA-08-142: Pharmacologic Agents and Drugs for Mental Disorders (SBIR [R43/R44]). Schizophrenia is a devastating brain disorder that affects approximately 0.7% of the global population. The symptoms of schizophrenia can be grouped into three major categories: positive (e.g. hallucinations and delusions), negative (e.g. social isolation and inappropriate emotional response),and cognitive symptoms. The cognitive symptoms of schizophrenia include impaired attention and disruption of working memory, an essential type of short term memory. These fundamental cognitive processes are critical for the performance of day-to-day activities, and their disruption in schizophrenia is a key factor underlying the inability of patients to integrate successfully into society. Indeed, the cognitive deficits associated with schizophrenia are recognized as a core component of the disorder. Currently available antipsychotic therapies are effective at ameliorating the positive symptoms of schizophrenia, but they are not effective at treating the negative or cognitive symptoms of the disease. Moreover, these therapies often cause significant side effects such as motor disturbances, weight gain, and diabetes. For these reasons, schizophrenia represents a major area of unmet medical need which must be addressed by the discovery and development of new antipsychotic therapies that are effective at treating the cognitive symptoms of schizophrenia with reduced potential for side effects. The overall goal of this proposal is to optimize a series of lead compounds we have identified with dual 5-HT2C receptor agonist and 5-HT6 receptor antagonist activities. Compounds with this novel pharmacological profile hold promise for yielding safe and effective treatments for the positive as well as the cognitive symptoms of schizophrenia. Such compounds are likely to have a significantly reduced risk for generating the undesirable side effects that are typically observed with currently available antipsychotic therapies. Successful completion of the specific aims outlined in this proposal will result in the identification of lead compounds with desirable pharmacological and drug-like properties. Furthermore, proof-of-concept for this novel target profile will be achieved by demonstrating antipsychotic and pro-cognitive efficacy in animal models with a selected lead compound. These achievements will provide a solid basis fora Phase II SBIR application to support further research aimed at selecting a clinical development candidate for the treatment of schizophrenia and its cognitive symptoms. A clinical development candidate with this novel profile would comprise a valuable asset within the pharmaceutical industry and would provide the basis of a successful commercial outcome for the research program. The research outlined in this proposal will directly benefit patients and their families and caregivers by leading to the discovery of safer and more effective medicines for the treatment of schizophrenia. PUBLIC HEALTH RELEVANCE: We have discovered a series of compounds with a unique target profile for specific serotonin receptors that are relevant to schizophrenia and cognition. Based on this discovery, we are optimizing these compounds to identify a new class of candidate drugs for treatment of schizophrenia and its severe cognitive deficits. Our goal is to discover new therapies that are more effective than current treatments with reduced potential for side effects. This research will thus have a positive impact on the significant population of schizophrenia patients and their family members and caregivers, which includes many millions of people worldwide. In particular,improving the cognitive outcome in schizophrenia will help patients to improve their job performance and to integrate into society more effectively.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 628.88K | Year: 2011
DESCRIPTION (provided by applicant): Schizophrenia is a debilitating disease that afflicts 1% of the US population. Symptoms of schizophrenia are classified into three categories: positive (delusions and hallucinations), negative (flattened affect, socialisolation) and cognitive (impaired attention, executive function, working memory). Existing antipsychotics, while effective at treating the positive symptoms, do not treat the negative or cognitive symptoms of schizophrenia making them a significant unmetmedical need. One of the challenges in discovering effective therapies for CNS diseases is development of preclinical assays that accurately model human cognitive processes. Traditional cell-based functional assays that model a specific target molecule orpathway implicated in a disease do not recapitulate the complex cellular and network processes underlying cognition. We propose the development of a novel in vitro brain slice screening platform that exploits the conservation of brain circuitry and functionality from rodents to humans. For example, human functional imaging and electroencephalographic studies have shown that higher cognitive functions, such as attention and working memory are associated with synchronized neuronal oscillations in the prefrontal cortex (PFC), particularly in the gamma frequency range (30-90Hz). Many of the cognitive impairments associated with schizophrenia are accompanied by a disruption of neural oscillations in the PFC. We have established an in vivo electrophysiological recording assay in freely behaving mice and have observed a significant alteration of gamma oscillations in the PFC of a genetic (calcineurin knockout) model of schizophrenia during the performance of attentional and mnemonic tasks. Our observations indicatethat neural oscillations recorded from the PFC of rodents in vivo can provide a robust and objective measure of cognition. As a first step towards modeling the complex neural activity associated with cognition in an in vitro assay, we have begun to develophigher throughput in vitro electrophysiological methods based on multielectrode array technology. Using acute brain slices that preserve adult network connectivity, we can recapitulate the same high-frequency synaptic activity associated with cognitive processing in vivo. Our preliminary data indicate that alterations in gamma oscillations observed in vivo are also observed in vitro, suggesting good feasibility in translating in vivo electrophysiological endophenotypes into in vitro screening assays. Thesein vitro assays will permit testing of compound efficacy on processes important for cognition at much earlier stages of development, mitigating many of the risks associated with preclinical in vivo pharmacology. Moreover, in vitro assays provide the higher bandwidth required for optimization of early stage compounds. This proposal will 1) characterize neural oscillations in vitro in different mouse models of schizophrenia and 2) validate these new in vitro screening paradigms using well studied reference compounds in use for treatment of the positive or cognitive symptoms of schizophrenia. PUBLIC HEALTH RELEVANCE: Schizophrenia is a debilitating mental illness that affects approximately 1% of the population worldwide. The research outlined in this proposal will support the development of innovative approaches and technologies to identify new therapies for the cognitive deficits in schizophrenia, which are a major unmet medical need. This research will have a positive impact on a significant populationof schizophrenia patients, their family members and caregivers. In particular, improving the cognitive outcome in patients will enhance their quality of life, enabling them to function more effectively in their personal and particularly their professionallives. This work is especially important, as the costs of unemployment due to schizophrenia have been estimated at 22 billion annually.
Galenea Corporation | Date: 2010-04-23
The present invention provides, in some aspects, methods for identifying agents useful in treating disorders or conditions associated with cognitive deficits. In some aspects, the invention provides methods for detecting a cognitive deficit in a subject.
Galenea Corporation | Date: 2014-11-04
Indole compounds are disclosed. Also disclosed are methods for using the compounds to treat human and animal disease, pharmaceutical compositions of the compounds, and kits including the compounds.
Galenea Corporation and United States Of America | Date: 2015-09-25
The present invention provides, in some aspects, methods for identifying and evaluating signatures in electroencephalographic oscillations that occur during onset of an exploratory activity in a subject.